Fig. 2. NCAM is partially localized in early endosomes. Cortical neurons isolated from E15.5 mice or B35 cells were transiently co-transfected with GFP-Rab5 and human NCAM140 or NCAM180. NCAM endocytosis was induced with an NCAM-specific antibody for 30 (cortical neurons) or 60 minutes (B35 cells) and cells were directly processed for fluorescence analysis of NCAM with GFP-Rab5. Photos were taken either from entire cells of cortical neurons or B35 cells (A) or from neurites and growth cones of cortical neurons (B). The experiment was carried out three times with at least 15 cells analysed in each experiment.